Testing for high‐risk HPV in cervical and tonsillar paraffin‐embedded tissue using a cartridge‐based assay

This study evaluates the suitability of Xpert HPV (Cepheid, Sunnyvale, CA, USA) test for cervical and tonsillar formalin‐fixed paraffin‐embedded (FFPE) tissue samples as compared to the tests currently used in diagnostics. Cervical biopsies and liquid cytology (LC) samples were collected from 48 women attending colposcopy. Biopsies were processed for histology and tested for hrHPV using Xpert HPV. LC samples were tested using Xpert and Hybrid Capture 2 (HC2; Qiagen, Hilden, Germany) tests. Also 29 archived tonsillar carcinoma samples were tested using Xpert, and the results were compared with histology and immunohistochemical p16INK4a (p16) staining. Among valid cervical LC samples 46.8% were hrHPV positive using Xpert test and 55.3% with HC2. The sensitivity of Xpert was 84.6% as compared to HC2, and overall test concordance was 91.5%. Test concordance between valid Xpert results from biopsies and LC samples was 84.6%. Among valid tonsillar samples 70.4% were hrHPV positive, and concordance of 96.3% was found between Xpert and p16 staining. To conclude, Xpert HPV test cartridge provides a convenient platform to test individual samples, including FFPE samples. Further studies are needed to establish whether test sensitivity is sufficient to reliably differentiate between hrHPV positive and hrHPV negative head and neck carcinomas.     

Comparison of Cepheid® Xpert Flu and Roche RealTime Ready Influenza A/H1N1 Detection Set for detection of influenza A/H1N1

Objective

To compare two influenza polymerase chain reaction (PCR) methods.

339例疑似肺结核患者Xpert Mtb/RIF检测临床分析

目的：探讨基层医院对结核病快速检测的方法。方法：收集2014年全年结核病门诊入选国家免费项目患者的痰标本,进行Xpert Mtb/RIF检测、荧光显微镜涂片找抗酸杆菌,并将二者进行对比,以培养做为最终标准。对结果进行分析。结果：对339份标本进行Xpert Mtb/RIF,荧光镜抗酸染色及培养,其中检测组175例患者标本中,阳性标本123例,阳性率为70.3%;对照组164中涂片阳性标本58例,阳性率为35.4%。卡方检验结果表明两组有显著性差异（P<0.01）。针对两种方法的阳性标本进行固体培养,培养结果为：Xpert Mtb/RIF检测阳性123例标本,阳性为80例,阳性率为70.3%。对照组阳性标本58例,培养出细菌19例,阳性率为19.3%卡方检验结果表明两组显著性差异（P<0.01）。结论：Xpert Mtb/RIF方法能快速、方便、灵敏检测结核杆菌,并快速培养出结核杆菌及耐药分析。     

Xpert MTB/RIF在骨关节结核患者

【摘要】 目的：探讨Xpert MTB/RIF在骨关节结核患者快速诊断中的应用价值。方法：2014年2月~2014年11月使用Xpert MTB/RIF对49例骨关节结核患者及32例非结核性骨关节病患者的脓液标本进行检测,以临床诊断为金标准,计算Xpert MTB/RIF检测结核分枝杆菌的敏感性、特异性、阳性预测值、阴性预测值、一致率;同时对脓液标本行抗酸染色及结核分枝杆菌快速培养（BACTECT MGIT 960）,比较Xpert MTB/RIF与抗酸染色、结核分枝杆菌快速培养在敏感性及特异性上的差异;综合上述两方面因素评价Xpert MTB/RIF在骨关节结核患者快速诊断中的应用价值。结果：Xpert MTB/RIF检测单个脓液标本的时间为2.3±0.2h。49例骨关节结核患者脓液标本中,46例Xpert MTB/RIF检测结核分枝杆菌阳性,3例Xpert MTB/RIF阴性;32例非结核性骨关节病患者脓液标本中,1例Xpert MTB/RIF检测结核分枝杆菌阳性,31例Xpert MTB/RIF阴性;以临床诊断为金标准,Xpert MTB/RIF检测结核分枝杆菌的敏感性为93.87%、特异性为96.87%、阳性预测值为97.87%、阴性预测值为91.17%,一致率为95.06%。在46例Xpert MTB/RIF结核分枝杆菌阳性的骨关节结核患者中,10例存在利福平耐药突变基因,耐药突变率为21.73%。49例骨关节结核患者脓液标本中,抗酸染色阳性8例,阴性41例,敏感性为17.39%;结核分枝杆菌快速培养阳性患者11例,阴性38例,敏感性为23.91%;32例非结核性骨关节病患者脓液标本抗酸染色及结核分枝杆菌快速培养均为阴性,其特异性均为100%。Xpert MTB/RIF检测结核分枝杆菌的的敏感性优于抗酸染色及结核分枝杆菌快速培养（P<0.05）,特异性与抗酸染色及结核分枝杆菌快速培养无明显差异（P>0.05）。结论：Xpert MTB/RIF在骨关节结核患者的快速诊断中具有较高的诊断效能,其耗时短,敏感性高,特异性强,与抗酸染色及结核分枝杆菌快速培养比较具有明显优势。     

结核感染T细胞斑点试验联合胸腔积液Xpert MTB/RIF检测诊断结核性胸腔积液的价值

目的评估结核感染T细胞斑点试验（T-SPOT.TB）联合胸腔积液结核分枝杆菌/利福平耐药实时荧光定量核酸扩增检测技术(Xpert MTB/RIF) 在结核性胸腔积液的诊断价值。方法收集胸腔积液患者112例,其中确诊或临床诊断为结核性胸腔积液76例,非结核性胸腔积液36例。所纳入患者治疗前均行胸腔积液T-SPOT.TB、血T-SPOT.TB、胸腔积液Xpert MTB/RIF、胸腔积液腺苷脱氨酶（ADA）检测,并对其检测结果进行分析。结果胸腔积液T-SPOT.TB诊断结核性胸腔积液的敏感度（97.37%,74/76),高于外周血T-SPOT.TB （76.32%,58/76）、胸腔积液Xpert MTB/RIF（65.79%,50/76）和胸腔积液ADA（28.95%,22/76）,差异均有统计学意义（χ2=14.74、25.22、76.45,P<0.01）。胸腔积液Xpert MTB/RIF检测的特异度（100%,36/36）均高于胸腔积液T-SPOT.TB（77.78%,28/36）、外周血T-SPOT.TB（55.56%,20/36）和胸腔积液ADA（50.00%, 18/36）,差异均有统计学意义（χ2=6.89、20.57、24.00,P均＜0.01）。胸腔积液T-SPOT.TB和胸腔积液Xpert MTB/RIF联合检测的敏感度（64.47%, 49/76）低于胸腔积液T-SPOT.TB单独检测（97.37%,74/76）,差异有统计学意义（χ2=26.63,P<0.01）,但特异度由77.78%(28/36)提高至100%(36/36),差异有统计学意义（χ2=45.82,P<0.01）。结论胸腔积液T-SPOT.TB检测结核性胸腔积液具有较高的敏感度和特异度,胸腔积液T-SPOT.TB和胸腔积液Xpert Mtb/ＲIF联合检测可明显提高诊断的特异度。     

Xpert MTB/RIF和T-SPOT.TB对涂阴肺结核诊断价值的评估

目的评估并比较Xpert MTB/RIF和T-SPOT.TB对涂阴肺结核的诊断价值。方法收集2014年12月至2015年12月天津市海河医院门诊和住院的呼吸疾病患者208例,分为结核组125例和对照组83例,同时对其痰液样本行涂片镜检、液体快速培养和Xpert MTB/RIF检测,对其血液样本行T-SPOT.TB试验。结果 Xpert MTB/RIF诊断涂阴肺结核的敏感性、特异性、ROC曲线下面积(AUC)及优势比(odds ratio,OR)分别为56.5%(95%CI:45.8%~66.8%)、98.8%(95%CI:93.5%~100.0%)、0.777(95%CI:0.708~0.836)及106.60(χ~2=62.87,P0.01);T-SPOT.TB诊断涂阴肺结核的敏感性、特异性、AUC及OR值分别为83.7%(95%CI:74.5%~90.5%)、62.7%(95%CI:51.3%~73.0%)、0.732(95%CI:0.660~0.796)及8.61(χ~2=39.44,P0.01);两者敏感性(χ~2=18.58)和特异性(χ~2=28.03)的差异有统计学意义(P0.05),AUC的差异无统计学意义(Z=1.235,P0.05);Xpert MTB/RIF阴性的涂阴肺结核患者T-SPOT.TB阳性率为70.0%。结论 Xpert MTB/RIF和T-SPOT.TB均有助于早期、快速、准确地辅助诊断肺结核,T-SPOT.TB可以作为Xpert MTB/RIF的后续试验辅助诊断Xpert MTB/RIF阴性的涂阴肺结核。     

Pattern of missing probes in rifampicin resistant TB by Xpert MTB/RIF assay at a tertiary care centre in Mumbai

Setting

Department of Microbiology.

Xpert HIV-1病毒载量检测系统用于HIV-1病毒载量检测的临床价值

目的探讨Xpert HIV-1病毒载量(Xpert HIV-1 VL)检测用于HIV-1病毒载量检测的临床价值。方法采用Xpert HIV-1 VL和Abbott m2000 Real Time检测系统(Abbott m2000)同时检测191份血浆样本的HIV-1病毒载量。采用符合率、卡方检验和Kappa值等统计分析两种检测方法定性检测结果的一致性,采用相关性、回归分析和Bland-Altman模型等统计分析两种检测方法定量检测结果的一致性。结果两种检测方法的阳性符合率、阴性符合率和总符合率均为100%,两种方法检测结果一致性分析的Kappa值为1.00(P<0.001);两种方法检测结果的相关系数为0.9576,回归分析表现出较高的相关性(R2=0.9170),Bland-Altman模型分析显示两种方法检测有95.3%(164/172)的样本在95%一致性界限内。结论Xpert HIV-1 VL用于检测HIV-1 RNA具有很高的灵敏度、准确性和稳定性,且操作简单、方便、安全,适合临床用于对HIV-1患者血浆样本中HIV-1 RNA的检测。     

Assessing the utility of the Xpert Mycobacterium tuberculosis/rifampin assay for analysis of bronchoalveolar lavage fluid in patients with suspected pulmonary tuberculosis

BackgroundThere is limited research assessing the utility of the Xpert Mycobacterium tuberculosis/rifampin (MTB/RIF) assay for the analysis of bronchoalveolar lavage fluid (BALF) in Chinese patients with suspected pulmonary tuberculosis (PTB). Thus, our objective was to determine the diagnostic accuracy of the Xpert MTB/RIF assay and evaluate its utility for the determination of rifampicin resistance.MethodsWe retrospectively analyzed BALF from 214 patients with suspected PTB between January 2018 and March 2019. Using mycobacterial culture or final clinical diagnosis as the reference standard, the diagnostic accuracy of the smear microscopy (SM), tuberculosis bacillus DNA (TB‐DNA), Xpert MTB/RIF assay, and the determination of rifampicin resistance based on the Xpert MTB/RIF assay were compared.ResultsAs compared to mycobacterial culture, the sensitivity of the Xpert MTB/RIF assay, SM, and TB‐DNA were 85.5% (74.2%–93.1%), 38.7% (26.6%–51.9%), and 67.7% (54.7%–79.1%), respectively. As compared to the final diagnosis, the specificity of the Xpert MTB/RIF assay, SM, and TB‐DNA were 100.0% (95.9%–100.0%), 94.3% (87.1%–98.1%), and 98.9% (93.8%–100.0%), respectively. The sensitivity and specificity of the rifampicin resistance detection using the Xpert MTB/RIF assay were 100% and 98.0%, respectively, with liquid culture as the reference.ConclusionsThis study demonstrates that the analysis of BALF with the Xpert MTB/RIF assay provides a rapid and accurate tool for the early diagnosis of PTB. The accuracy of diagnosis was superior compared with the SM and TB‐DNA. Moreover, Xpert is a quick and accurate method for the diagnosis of rifampicin‐resistant tuberculosis and can also provide more effective guidance for the treatment of PTB or multidrug‐resistant tuberculosis (MDR‐TB).